Protocols

This screen is designed to identify genes regulating IgE production in vivo in response to aluminum hydroxide (alum)-emulsified ovalbumin (OVA) [OVA-alum]. Alum is an adjuvent usied in human vaccines. Mice are immunized on day 0, without any boosting step on day 14, and serum is collected and analyzed for antigen-specific IgE on day 14. ENU-mutatgenized G3 mice that produce a increased amount of OVA-specific IgE(normalized by the level of the total IgE), relative to wild type mice are identified as potential mutants.

Dilute the OVA in 0.9% saline to 1000ug/ml to make the OVA-Alum solution (below)

A solution of 10 ug/mL OVA in PBS will be needed for the ELISA

OVA-Alum solution

*** Always make OVA solution fresh the day of the injection.***

1. Mix equal parts the OVA solution and Alydrogel 2%. For example, for one mouse mix 100 μL OVA solution and 100 μL of the Alhydrogel 2% together. Mix well by pipetting up and down for at least 5 minutes to allow Alhydrogel 2% to effectively adsorb the antigen.